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Aquaculture is a sector with a rapid growth and a great potential to provide healthy and sustainable protein for future generations. In this context, and with stricter criteria of farmed animal welfare, it is very important to monitor fish behavior for the simultaneous improvement of the productivity and the welfare of group-housed animals. In the framework of AQUAEXCEL2020 EU project we have developed an ultra-low-power, stand-alone, miniature microsystem (AEFishBIT) with the double aim to combine measurements of physical and metabolic activities. The system was developed using available technology and includes a triaxial accelerometer and a microcontroller packaged in a waterproof container. It has been designed to be externally implanted on fish operculum and by signal filtering, it allows the measurements of physical activity and respiratory frequency., This project has received funding from the European under grant agreement No. 652831 (AQUAEXCEL2020, Aquaculture infrastructures for excellence in European fish view and the European Union cannot be held responsible for any use that may be made of the information contained herein. Additional funding was obtained from projects SURF (TEC2014-60527-C2-1-R), FICASES (TEC2017-89403-C2-2-R), from of the Spanish Ministry of Economy and Competitiveness, and ProID2017010062, from Canarian Agency for Research, Innovation and Information Society (Gobierno de Canarias), all of them co-funded European Regional Development Funds (FEDER). We acknowledge the support from EnABLES European project, and in particular from Dr. Michael Hayes and Prateek Asthana, from Tyndall National Institute, Cork, Ireland.

[EN] The invention relates to a device placed on the gill cover to monitor the metabolic state and well-being of fish through joint, simultaneous measurements of breathing frequency and physical activity via a plurality of accelerations that are coplanar (x and y axes) and normal to the gill cover (z axis). The substrate is flexible and is covered with an impervious coating, which contains an electronic circuit with an acceleration sensor (208), a control unit (202), a microcontroller (206) configured to execute at least one activity-monitoring routine, a memory (210) and a battery (204). Data are collected via an external unit (201) which can optionally process them in a complete or partial manner. This type of measurement can be processed jointly with another type of measurement, obtained from other sensors of environmental parameters, [ES] Dispositivo emplazado en el opérculo para supervisar el estado metabólico y bienestar de peces a través de medidas conjuntas y simultáneas de frecuencia respiratoria y actividad física mediante una pluralidad de aceleraciones coplanares (ejes x,y) y normales al opérculo (eje z). El sustrato es flexible y está cubierto con un revestimiento impermeable, donde se encuentra un circuito electrónico con sensor de aceleración (208), una unidad de control (202), un microcontrolador (206) configurado para ejecutar al menos una rutina de monitorización de actividad, una memoria (210) y una batería (204). La toma de datos se efectúa a través de una unidad externa (201) que eventualmente puede procesarlos de forma total o parcial. Este tipo de mediciones se pueden procesar de forma conjunta con otro tipo de medidas, obtenidas a partir de otros sensores de parámetros medio-ambientales, [FR] La présente invention concerne un dispositif mis en place sur l'opercule pour surveiller l'état métabolique et le bien-être de poissons au moyen de mesures conjointes et simultanées de la fréquence respiratoire et de l'activité physique avec une pluralité d'accélérations coplanaires (axes x,y) et perpendiculaires à l'opercule (axe z). Le substrat est flexible et couvert d'un revêtement imperméable, où se situe un circuit électronique avec capteur d'accélération (208), une unité de commande (202), un microcontrôleur (206) configuré pour exécuter au moins une routine de surveillance d'activité, une mémoire (210) et une batterie (204). La capture de données est effectuée au moyen d'une unité externe (201) qui peut éventuellement les traiter de manière complète ou partielle. Les mesures de ce type peuvent être traitées de manière conjointe avec un autre type de mesures, obtenues à partir d'autres capteurs de paramètres du milieu ambiant, Consejo Superior de Investigaciones Científicas (España), Universidad de Las Palmas de Gran Canaria, A1 Solicitud de patente con informe sobre el estado de la técnica

Biosensor technology for tracking individual challenged fish behaviour has the potential to revolutionize aquaculture, allowing farmers and breeders to orientate selective breeding towards more robust and efficient fish or improve culture conditions for a more sustainable and ethical production. The proposed solution within the AQUAEXCEL2020 EU project is a stand-alone, small and light (1 g) device (AEFishBIT), based on a tri-axial accelerometer and a microprocessor. It is externally attached to the operculum to monitor physical activity by mapping accelerations in x- and y-axes, while operculum beats (z-axis) serve as a measurement of respiratory frequency. The conducted operculum attachment protocol does not show signs of tissue damage or growth impairment in active feeding gilthead sea bream. AEFishBIT offers a wide range of new information based on individual behaviour, allowing to point out the asynchrony of movements as an indirect measure of aging and adaptability to farming environment, as well as to discriminate different coping behaviour (proactive or reactive) of gilthead sea bream challenged with low water oxygen concentrations. AEFishBIT also provides reliable information of disease outcome in fish parasitized with an intestinal myxozoan, emerging as a powerful tool for sensing the quality of the environment and improving selective breeding protocols., The study has received funding from the European Union’s Horizon 2020 research and innovation programme, GA no 652831 (AQUAEXCEL2020).

Dispositivo emplazado en el opérculo para supervisar el estado metabólico y bienestar de peces a través de medidas conjuntas y simultáneas de frecuencia respiratoria y actividad física mediante una pluralidad de aceleraciones coplanares (ejes x,y) y normales al opérculo (eje z). El sustrato es flexible y está cubierto con un revestimiento impermeable, donde se encuentra un circuito electrónico con sensor de aceleración (208), una unidad de control (202), un microcontrolador (206) configurado para ejecutar al menos una rutina de monitorización de actividad, una memoria (210) y una batería (204). La toma de datos se efectúa a través de una unidad externa (201) que eventualmente puede procesarlos de forma total o parcial. Este tipo de mediciones se pueden procesar de forma conjunta con otro tipo de medidas, obtenidas a partir de otros sensores de parámetros medio-ambientales, Consejo Superior de Investigaciones Científicas (España), Universidad de Las Palmas de Gran Canaria, A1 Solicitud de patente con informe sobre el estado de la técnica

Dispositivo emplazado en el opérculo para supervisar el estado metabólico y bienestar de peces a través de medidas conjuntas y simultáneas de frecuencia respiratoria y actividad física mediante una pluralidad de aceleraciones coplanares (ejes x,y) y normales al opérculo (eje z). El sustrato es flexible y está cubierto con un revestimiento impermeable, donde se encuentra un circuito electrónico con sensor de aceleración (208), una unidad de control (202), un microcontrolador (206) configurado para ejecutar al menos una rutina de monitorización de actividad, una memoria (210) y una batería (204). La toma de datos se efectúa a través de una unidad externa (201) que eventualmente puede procesarlos de forma total o parcial. Este tipo de mediciones se pueden procesar de forma conjunta con otro tipo de medidas, obtenidas a partir de otros sensores de parámetros medio-ambientales, Consejo Superior de Investigaciones Científicas (España), Universidad de Las Palmas de Gran Canaria, B2 Patente con examen previo

Integration of technological solutions aims to improve accuracy, precision and repeatability in farming operations, and biosensor devices are increasingly used for understanding basic biology during livestock production. The aim of this study was to design and validate a miniaturized tri-axial accelerometer for non-invasive monitoring of farmed fish with re-programmable schedule protocols. The current device (AE-FishBIT v.1s) is a small (14 mm × 7 mm × 7 mm), stand-alone system with a total mass of 600 mg, which allows monitoring animals from 30 to 35 g onwards. The device was attached to the operculum of gilthead sea bream (Sparus aurata) and European sea bass (Dicentrarchus labrax) juveniles for monitoring their physical activity by measurements of movement accelerations in x- and y-axes, while records of operculum beats (z-axis) served as a measurement of respiratory frequency. Data post-processing of exercised fish in swimming test chambers revealed an exponential increase of fish accelerations with the increase of fish speed from 1 body-length to 4 body-lengths per second, while a close relationship between oxygen consumption (MO2) and opercular frequency was consistently found. Preliminary tests in free-swimming fish kept in rearing tanks also showed that device data recording was able to detect changes in daily fish activity. The usefulness of low computational load for data pre-processing with on-board algorithms was verified from low to submaximal exercise, increasing this procedure the autonomy of the system up to 6 h of data recording with different programmable schedules. Visual observations regarding tissue damage, feeding behavior and circulating levels of stress markers (cortisol, glucose, and lactate) did not reveal at short term a negative impact of device tagging. Reduced plasma levels of triglycerides revealed a transient inhibition of feed intake in small fish (sea bream 50–90 g, sea bass 100–200 g), but this disturbance was not detected in larger fish. All this considered together is the proof of concept that miniaturized devices are suitable for non-invasive and reliable metabolic phenotyping of farmed fish to improve their overall performance and welfare. Further work is underway for improving the attachment procedure and the full device packaging., This project has received funding from the European Union’s Horizon 2020 Research and Innovation Programme under grant agreement No. 652831 (AQUAEXCEL2020, Aquaculture infrastructures for excellence in European fish research towards 2020). This publication reflects the views only of the authors, and the European Commission cannot be held responsible for any use which may be made of the information contained therein. Additional funding was obtained from project SURF (TEC2014-60527-C2-1-R) of the Spanish Ministry of Economy and Competitiveness and the European Regional Development Fund (FEDER).

La importancia de determinar el sexo en especies monomórficas que viven en cautiverio conlleva a estrategias de apareamiento a fin de poder incentivar la reproducción. Existen métodos no moleculares de reconocimiento pero son factibles en edad adulta y mucho más difícil cuando son polluelos. Mediante la técnica molecular de PCR, se amplificó el gen de la Cromo Helicasa de Unión al ADN empleando los cebadores, 2550F/2718R. Por este método molecular se consiguió determinar el sexo en 19 de 23 especies de las cuales 3 son ejemplares de: Amazona aestiva, 1 de Pipile grayi, 1 de Bubo virginianus, 4 de Ara chloropterus, 6 de Crax fasciolata, 2 de Caracara plancus, 3 de Chauna torquata, 1 de Cariama cristata y 2 de Cairina moschata del Centro de Investigación de Animales Silvestres de ITAIPU binacional, del lado paraguayo. Nuestros resultados sugieren que el par de cebadores 2550F/2718R podría ser de utilidad para determinar el sexo en las especies estudiadas en el país

Trabajo presentado en Aquaculture Europe 2019, celebrado en Berlín (Alemania), del 7 al 10 de octubre de 2019, The use of new technologies for individual and non-invasive monitoring of farmed fish is becoming an urgent demand of the aquaculture industry (Føre et al., 2018). This offers the possibility of the measurement of a range of variables that are directly or indirectly related with metabolic condition, health and welfare status (Metcalfe et al., 2016; Neethirajan et al., 2017). The solution proposed within the AQUAEXCEL2020 EU project is a smart device named AEFishBIT, developed and validated to provide reliable and simultaneous measurements of physical activity and respiratory frequency. The device is attached in the operculum using a fairly invasive procedure, and initial testing has been conducted in gilthead sea bream and European sea bass, the two most important farmed fish of Mediterranean aquaculture. The AEFishBIT is a tri-axial accelerometer for recording and processing acceleration data from x-, y- and z-axes. Records of operculum breathing (z-axis acceleration) served as a direct measurement of respiratory frequency, whereas estimation of fishactivity was derived from the x- and y-axis signals. The device also includes a passive RFID tagging device for rapid identification.The finalweight of the full packaged device is less than 1 g in air. The autonomy of the system in stand-alone mode is 6 h of continuous data recording with different programmable time schedules. Algorithms to convert accelerometer measurements in physical activity and respiratory frequency (breaths/s) have been validated with exercised juveniles in a 10-L swim tunnel respirometer (Loligo® Systems). Fish were submitted to controlled speeds from 1 body-length per second (BL/s) until exhaustion by 0.5 BL/s increase steps, with measurements of O2 consumption rates (MO2). AEFishBIT was programmed for 2-min time window at different intervals. The close parallelism between the inferred respiratory frequency and MO2 highly validates the use of the proposed algorithm to infer respiratory frequency in exercised sea bream and sea bass (1-4 BL/s speed range). Likewise, the physical activity index increased exponentially rather than linearly with the increase of swimming speed, as the total jerk magnitude only reflects fish-related accelerations. Regarding fish species differences, it is important to note that, at the same swimming speed, MO2 and the calculated respiratory frequency were consistently higher in sea bream than in sea bass. AEFishBIT measurements in free-swimming fish evidenced different chronotypes for a given fish population, termed as ¿diurnal¿, ¿nocturnal¿ or ¿arrhythmic¿ on the basis of the moment of the day when the maximum activity pattern (light phase, dark phase or undifferentiated, respectively) is achieved. AEFishBIT measurements also offer the possibility to discriminate different personality traits when fish are challenged with aquaculture stressors (e.g. hypoxia, stocking density). The present work represents the proof of concept of a miniaturized device for individual fish phenotyping of metabolic condition and welfare. The different behavior patterns of free-swimming fish can be associated with a better performance or differences in stress and disease resilience. These monitored features also evoke changes in season, age, development or nutritional condition, remaining to establish the involvement of genetic and epigenetic factors on the measured parameters. This opens new research opportunities for the individual fish phenotyping of productive traits through the production cycle, but also for selective breeding in combination with other now promising omics approaches (e.g transcriptomics, methylomics, metabolomics, microbiomics)., The study has received funding from the European Union’s Horizon 2020 research and innovation programme under Grant Agreement no 652831 (AQUAEXCEL2020)

Trabajo presentado en el XII Conference of the Iberian Association for Comparative Endocrinology (AIEC), celebrado en Faro (Portugal), del 26 al 28 de septiembre de 2019, With the use of electronic tracking technology, data on fish movements, physiology and environmental parameters can be collected from each tagged individual. The perfect tag - small size, lasting activity, long distance data transmission (e.g. radiotransmitters, acoustic transmitters, pop-up satellite archival tags) - does not exist, and the choice of telemetry technology and tagging method (external, intracelomic, intragastric) depends on fish species, life stage and research question. The proposed solution within the AQUAEXCEL2020 EU project is a stand-alone, small and light (1 g) device (AEFishBIT), composed of a tri-axial accelerometer, a microprocessor, a battery and an RFID tag for quick-smart identification. The device is externally attached to the operculum to monitor physical activity by mapping accelerations in x- and y-axes, while operculum beats (z-axis) serve as a measurement of respiratory frequency. Initial functional validation has been made with gilthead sea bream and European sea bass juveniles in swimming test chambers, highlighting a high correlation of oxygen consumption and swimming activity with the calculated AEFishBIT records (MartosSitcha et al., 2019; Frontiers in Physiology 10:667). Current studies with free-swimming fish reveal that the attachment protocol does not have a negative impact on growth performance in active feeding fish, offering a wide range of new information based on individual behaviour: i) the switch from nocturnal to diurnal rhythms of activity across season, ii) the different energy partitioning for growth and maintenance, iii) the asynchrony of movements as an indirect measure of aging and adaptability to culture environment, and iv) the different coping behaviour of fish challenged with low water oxygen concentrations. AEFishBIT also provides reliable information of disease outcome in fish parasitized with the intestinal myxozoan Enteromyxum leei, emerging as the ideal complement for sensing the quality of the environment and improving selective breeding protocols., AQUAEXCEL2020 EU Project, GA 652831

RESUMEN En Paraguay, no se han registrado casos autóctonos de malaria desde el 2011. Se realizó un estudio descriptivo observacional transversal en 6 monos y 23 aves que vivían en una región históricamente endémica de Paraguay para buscar presencia de reservorios silvestres de parásitos plasmodios causantes de la malaria. El ADN se extrajo por el método de Chelex a partir de una gota de sangre en un papel de filtro, y la detección del parásito se realizó mediante la PCR múltiple semianidada. Por este método, no se detectaron parásitos en ninguna de las 29 muestras. Se evaluó el riesgo potencial de circulación selvática de los parásitos que causan la malaria. Teniendo en cuenta la presencia de mosquitos anofelinos vectores en la zona, el hecho de que no se hayan observado casos positivos es un buen indicador teniendo en cuenta que nuestro país fue declarado recientemente como país libre de malaria por la OMS. ABSTRACT In Paraguay, autochthonous cases of malaria have not been recorded since 2011. A cross-sectional observational descriptive study was conducted in 6 monkeys and 23 birds living in a historically endemic region of Paraguay to identify wild reservoirs of plasmodium parasites that cause malaria. DNA was extracted by the Chelex method from a blood drop in a filter paper, and parasite detection was performed by the seminested multiplex PCR. By this method, parasites were not detected in any of the 29 samples. The risk of potential sylvatic circulation of the parasites causing malaria was evaluated. Considering the presence of anopheline mosquitoes in the area, the fact that we did not find any positive cases is a good indicator as our country was recently certified as a malaria-free country by the WHO.